J. Environ. Res. Develop. Journal of Environmental Research And Development Vol. 8 No. 2, October-December 2013

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1 ASCORBATE EFFECT ON THE LEAD CHLORIDE INDU- CED ALTERATIONS IN THE PROTEIN CONTENTS OF FRESH WATER FISH, Channa orientalis (Schneider) Borane V. R.*, Shinde B.R. 1 and Zambare S.P Department of Zoology, Jijamata Arts, Sci. and Com. College, Nandurbar, Maharashtra (INDIA) 2. Department of Zoology, Dr. B. A. Marathwada University, Aurangabad, Maharashtra (INDIA) Received July 10, 2013 Accepted November 13, 2013 ABSTRACT Fresh water fishes, Channa orientalis were exposed to chronic dose of lead chloride without and with ascorbic acid. Protein content from liver and kidney were estimated. Remarkable decrease in protein was observed in lead exposed fishes as compared to control. Fishes exposed to lead along with L-ascorbic acid showed increases protein in tissues than those exposed to only heavy metal. Pre-exposed fishes to heavy metal showed fast recovery in protein with ascorbic acid than in normal water. Key Words : Channa orientalis, Ascorbic acid (50mg/l.), Lead, Protein content- liver, Kidney INTRODUCTION Contamination of freshwater with heavy metal such as heavy metal has become a matter of great concern over the past decades not only because of their threat to public water supplies but also because of the damage caused to aquatic life especially fishes. 1 Fish are widely used to evaluate the health of aquatic ecosystems because pollutants build up in the food chain and are responsible for adverse effects and death in the aquatic systems. 2 Toxicity testing of chemicals on animals has been used for a long- time to detect the potential hazards posed by chemicals to man. Bioassay technique has been the cornerstone of programmes on environmental health and chemical safety. 3 Routes of exposure to lead include contaminated air, water, soil, food and consumer products. Trace metals are biological interest because of their role as micronutrients enter in aquatic organisms through the skin, gills, alimentary canal and by feeding. Although lead poisoning is one of the oldest known work and environmental hazards, the modern understanding of the small amount of lead necessary to cause harm did not come about until the latter half of the 20th century. No safe threshold for lead exposure has been discovered *Author for correspondence 268 that is, there is no known amount of lead that is too small to cause the body harm. Lead poisoning (also known as plumbism, Colica Pictonum, Saturnism, Devon colic, or Painter's colic) is a medical condition caused by increased levels of lead in the body. Lead is ubiquitous environmental toxin interferes with a variety of body processes that induces physiological, biochemical process and is toxic to many organs and tissues including the heart, bones, intestines, kidneys, reproductive and nervous systems. 4 It is particularly toxic to children, causing potentially permanent learning and behavior disorders. Prevention of lead exposure can range from individual efforts (e.g. removing lead-containing items such as piping or blinds from the home) to nationwide policies (e.g. laws that ban lead in products or reduce allowable levels in water or soil). The major treatments are removal of the source of lead and chelation therapy (administration of agents that bind lead so it can be excreted). Proteins are essential organic constituent which play a vital role in the cellular metabolism. It has been reported that depletion in the protein content in intoxicated fishes may be due to the higher affinity of metal compounds toward different amino acid residues of protein or due to proteolytic activity. The ascorbate effect on methyl mercury toxicity in reproductive organs

2 of guinea pigs and found the recovery in the metabolic functions indicate positive role in detoxification studied. AIMS AND OBJECTIVES To study the role of ascorbic acid after the exposure to PbCl 2 for chronic concentration. MATERIAL AND METHODS Medium sized fresh water fishes, Channa orientalis were collected from Shiven river area Nandurbar Dist. Nandurbar, Maharashtra, India in Fishes were washed with 0 1% KMNO 4 solution to avoid dermal infection. The physicochemical parameters determined by the APHA, AWWA and WACF standard method. 5 The fishes were divided into three groups A, B and C. Group A fishes as a control. The Group B fishes were exposed to LC 50/10 dose of (2.867ppm) as lead chloride for 45 days, while group C fishes were exposed to chronic concentration of heavy metal with ascorbic acid for 45 days. Fishes from B groups were divided into two groups after 45 days exposure to lead into DandE groups. Fishes of D groups were allowed to cure naturally in normal water, while those of E groups were exposed to ascorbic acid.liver and kidney from A, B and C group fishes after 15, 30 and 45 days of exposure and from D and E groups after 5 and 10 days of recovery were removed and dried at 80 ºC in the oven. Proteins were estimated by Folin Phenol reagent. RESULTS AND DISCUSSION The data obtained regarding the physicochemical properties is given in Table 1 while the protein contents in different tissues after exposure to lead with and without ascorbic acid and during recovery are given in the Tables 2 to Table 3. After chronic exposures to PbCl 2, protein contents in the different tissues of Channa orientalis were found to be depleted. The changes in the protein contents of a tissue due to heavy metal stress indicate the changes in the activity of the organism. It reflects in the utilization of their biochemical energy to counteract the toxic stress. In the presence of ascorbic acid the protein depletion is less as compared to those of lead chloride intoxicated fishes. The fishes pre-exposed to heavy metals salts showed fast recovery in the protein level in the presence of ascorbic 269 acid than those allowed to cure naturally. Alterations in biochemical values in fish give an indication and help to understand the mode of action and type of pollutants. Proteins are important organic constituents in animals which play an important role in cellular metabolism. Decline in protein may be due to enhanced protein catabolism, lead causes depletion of protein bound sulfhydryls with an increased oxidative stress to cell. 6 The depletion in tissue proteins of Channa orientalis may be due to impairment or low rate of protein synthesis under metal ion stress or due to utilization in the formation of mucoproteins which are released in the form of mucous. 7 The decline in total protein content in different tissues was directly proportional to the concentration of cadmium and duration of exposure. 8 Protein serve as energy source during stress condition also showed decreasing trend in the tested organ, maximum depletion was observed in liver, muscle and gill followed by kidney and brain depletion was equal in liver and muscle. 9 Ascorbic acid is essential for normal development and in most species it is produced endogenously, resembling a hormone. Ascorbate can serve as both serve as both pro - oxidant and antioxidant and also cross over effect of catalytic metal concentration. In present study depletion of protein content in different tissues of lead treated animals due to alterations of membrane permeability. The recovery of protein contents on ascorbic acid plays an important role against lead toxicity Channa orientalis (Fig. 1 and Fig. 2). The effect of toxic metal can be protected by ascorbic acid not only confers protection against lead toxicity, but it can also perform therapeutic role. 10 The stimulatory action of ascorbic acid is indicated by increase in protein content, and level of lysosomal enzymes enhanced capacity for phagocytosis. 11 Ascorbic acid as one of the most important antioxidants spares the other oxidants forming the first line of defense against free radicals and peroxides that are generated during cellular metabolism. 12 Ascorbic acid reduces lead level in the body by decreasing the rate of intestinal absorption. 13 Ascorbic acid also complexes lead in vivo thus acting as potential chelating agent and seem to alleviate lead

3 induced vitamin deficiency and metabolic disturbances. 14 Lead poisoning in the kidney of 270 rats was prevented of by calcium and ascorbic acid supplementation Table 1 : Physico-chemical parameters of water used for experimentation Temperature 25 1 ± ph ± 0 3 Conductivity 140 ± 15.7 µ mho -cm. Free CO ±1 3ml -1 Dissolved O ± 1 1ml -1 Total hardness 204 ± 12 0mg -1 Total alkalinity ± 32.8 mg -1 Magnesium ± 2.9 mg -1 Calcium ± 3.06 mg -1 Chloride ± mg -1 Table 2 : Protein contents in the liver of Channa orientalis after chronic exposure to without and with ascorbic acid and during recovery Treatment 15d 30d 45d 50d 55d Control 68.01± ± ± (2.867 ppm) (2.867 ppm)+ AA Recovery in normal water Recovery in AA ± *** (-17.12) ±0.5013*** (- 4.18) ±0.7512*** (-18.61) ±0.7202*** (-13.43) ±0.5519*** (-23.25) ±0.4663** (-13.86) ±.7352 ΔΔ [+10.75] 55.51± ΔΔΔ [+17.06] ± ΔΔ [+12.58] 58.28± ΔΔΔ [+22.90] (Values represent % in dry wt.) Table 3 : Protein contents in the kidney of Channa orientalis after chronic exposure to without and with ascorbic acid and during recovery Treatment 15d 30d 45d 50d 55d Control 62.83± ± ± (2.867ppm) (2.867ppm)+ AA Recovery in Normal water Recovery in AA ± *** (-9.07) ± ** (-6.19) ± *** (-9.99) ± ** (-6.58) ± *** (-15.18) ± ** (-8.43) ± Δ [+5.71] ± ΔΔ [+9.85] ± ΔΔ [+9.27] ± ΔΔΔ [+14.00] Values represent % in dry wt., AA = Ascorbic acid (50 mg/l., ± indicates S.D. of three observations, Values in ( ) indicates percent change over respective control, Values in [ ] indicates percent change over 45 days of respective B. *indicates significance with the respective control, Δ indicates significance with 45 days of respective B, p<0.05 = * & Δ, p<0.01 = **& Δ Δ, p<0.001 = *** & Δ Δ Δ, NS and Δ NS = Not significant

4 Protein contents (mg/100mg) PbCl 2 PbCl 2+AA Days Fig. 1 : Protein contents in liver of Channa orientalis after chronic exposure to lead without and with ascorbic acid and during recovery (mg/100mg) Protein contents (mg/100mg) PbCl 2 PbCl 2+AA Days Fig. 2 : Protein contents in Kidney of Channa orientalis after chronic exposure to lead without and with ascorbic acid and during recovery (mg/100mg) CONCLUSION In the present study depletion of protein contents in liver than kidney of PbCl 2 treated animals due to alterations of membrane permeability also due to enhanced protein catabolism. The recovery of protein contents on ascorbic acid plays an important role as both pro - oxidant and antioxidant against PbCl 2 toxicity in Channa orientalis. Fast recovery in protein contents than those which were allowed to cure naturally. ACKNOWLEDGEMENT Authors are thankful to Western Regional Office (WRO, UGC), Pune for providing financial assistance in the form of MRP. REFERENCES 1. Tawari- Fufeyin P., Igetei J. and Okoidigun M.E., Changes in the catfish, (Clarius gariepinus) exposed to acute cadmium and lead poisoning, Biosci. Res. Commun., 20(5), , (2008). 2. Vinodini R. and Narayanan M. Bioaccumulation of heavy metals in organs of fresh water fish, Cyprinus carpio (common carp). Int. J. Environ. Sci. Tech., 5(2), , (2008). 3. Ward G.S. and Parrish P.R., Manual of methods in aquatic environmental research, Part 6. Toxi. Tests, FAO Fisheries Technical Paper, No 185 FIRI/T185, (1982). 271

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